Sexed Sperm Bulk Separation Systems

Abstract

A broad object of a the instant invention can be to provide a method for separating X and Y sperm cells within a sample sperm cell population, the method including (i) differentiating between and (ii) separating sperm cells that have undergone a cellular process and sperm cells that have not undergone the cellular process, whereby a majority of the sperm cells that have undergone the cellular process can comprise one of X or Y sperm cells, and a majority of the sperm cells that have not undergone the cellular process can comprise the other of X or Y sperm cells. As to particular embodiments, the cellular process can be a maturational step. As to particular embodiments, the maturational step can be capacitation. As to particular embodiments, the maturational step can be the acrosome reaction. As to particular embodiments, non-viable and viable sperm cells can also be (i) differentiated between and (ii) separated.

1 . A method for separating X chromosome-bearing sperm cells and Y chromosome-bearing sperm cells within a sample sperm cell population, comprising:
 differentiating between sperm cells that have undergone a cellular process and sperm cells that have not undergone said cellular process; and   separating said sperm cells that have undergone said cellular process and said sperm cells that have not undergone said cellular process;   wherein a majority of said sperm cells that have undergone said cellular process comprise one of said X chromosome-bearing sperm cells or said Y chromosome-bearing sperm cells; and   wherein a majority of said sperm cells that have not undergone said cellular process comprise the other of said X chromosome-bearing sperm cells or said Y chromosome-bearing sperm cells.   
     
     
         2 . (canceled) 
     
     
         3 . The method of  claim 1 , further comprising:
 differentiating between sperm cells that have undergone a maturational step and sperm cells that have not undergone said maturational step; and   separating said sperm cells that have undergone said maturational step and said sperm cells that have not undergone said maturational step;   wherein a majority of said sperm cells that have undergone said maturational step comprise one of said X chromosome-bearing sperm cells or said Y chromosome-bearing sperm cells; and   wherein a majority of said sperm cells that have not undergone said maturational step comprise the other of said X chromosome-bearing sperm cells or said Y chromosome-bearing sperm cells.   
     
     
         4 . The method of  claim 3 , wherein said maturational step comprises capacitation; and
 wherein said differentiating between sperm cells that have undergone said capacitation and sperm cells that have not undergone said capacitation is facilitated by Y chromosome-bearing sperm cells undergoing said capacitation more quickly than X chromosome-bearing sperm cells.   
     
     
         5 - 6 . (canceled) 
     
     
         7 . The method of  claim 4 , further comprising inducing said capacitation to generate a capacitation-induced sperm cell subpopulation in which said capacitation is induced in at least a portion of said Y chromosome-bearing sperm cells. 
     
     
         8 - 20 . (canceled) 
     
     
         21 . The method of  claim 7 , further comprising employing a capacitation indicator which is associated with said capacitated sperm cells to differentiate said capacitated sperm cells from said non-capacitated sperm cells. 
     
     
         22 . The method of  claim 21 , furthering comprising associating a capacitation indicator associator with said capacitation indicator to differentiate said capacitated sperm cells from said non-capacitated sperm cells. 
     
     
         23 . The method of  claim 22 , wherein said capacitation indicator comprises a saccharide. 
     
     
         24 . (canceled) 
     
     
         25 . The method of  claim 23 , wherein said capacitation indicator associator comprises a lectin which associates with said saccharide. 
     
     
         26 - 39 . (canceled) 
     
     
         40 . The method of  claim 22 , further comprising differentiating between sperm cells that have undergone the acrosome reaction and sperm cells that have not undergone said acrosome reaction;
 wherein said sperm cells that have undergone said acrosome reaction comprise acrosome-reacted sperm cells; and   wherein said sperm cells that have not undergone said acrosome reaction comprise non-acrosome-reacted sperm cells.   
     
     
         41 . The method of  claim 40 , further comprising employing an acrosome reaction indicator which is associated with said acrosome-reacted sperm cells to differentiate said acrosome-reacted sperm cells from said non-acrosome-reacted sperm cells. 
     
     
         42 . The method of  claim 41 , furthering comprising associating an acrosome reaction indicator associator with said acrosome reaction indicator to differentiate said acrosome-reacted sperm cells from said non-acrosome-reacted sperm cells. 
     
     
         43 . The method of  claim 42 , wherein said acrosome reaction indicator comprises equatorin. 
     
     
         44 . (canceled) 
     
     
         45 . The method of  claim 43 , wherein said acrosome reaction indicator associator comprises an antibody which associates with said equatorin. 
     
     
         46 - 47 . (canceled) 
     
     
         48 . The method of  claim 42 , further comprising a first particle associated with said capacitation indicator associator, said first particle separable from said sample sperm cell population. 
     
     
         49 . The method of  claim 48 , wherein said first particle is separable from said sample sperm cell population by the application of a force to which said first particle is responsive. 
     
     
         50 . The method of  claim 49 , further comprising applying said force to said sample sperm cell population. 
     
     
         51 . The method of  claim 50 , wherein said force comprises a centrifugal force, an electrostatic force, a gravitational force, a magnetic force, or the like, or combinations thereof. 
     
     
         52 - 60 . (canceled) 
     
     
         61 . The method of  claim 49 , wherein said capacitation indicator associator is conjugated to said first particle to provide a capacitation indicator associator particle. 
     
     
         62 - 79 . (canceled) 
     
     
         80 . The method of  claim 61 , further comprising second particle associated with said acrosome reaction indicator associator, said second particle separable from said sample sperm cell population:
 wherein said acrosome reaction indicator associator is conjugated to said second particle to provide an acrosome reaction indicator associator particle.   
     
     
         81 - 97 . (canceled) 
     
     
         98 . The method of  claim 80 , further comprising:
 differentiating between non-viable sperm cells and viable sperm cells; and   separating said non-viable sperm cells and said viable sperm cells.   
     
     
         99 - 128 . (canceled)